Effect of storage period of the culture medium (M16) on the development of mouse zygotes in vitro and in vivo

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The present study was undertaken to examine the effect of storage period of the culture medium, M16 (Table 1), on the development of mouse zygotes to blastocyst stage in vitro, and to live fetuses after transfer to recipients.<BR>Mouse zygotes obtained from the superovulated F1 (C57BL/6×CBA) females 18 hrs after hCG injection were used. The zygotes with two pronuclei were cultured for 4 days in one of the medium either fresh, storaged for 30 days, 60 days, or 90 days in the refrigerator. Blastocysts developed from zygotes cultured in each medium were separately transferred into the uteri of recipients on the third or the 4th day of psedopregnancy. They were killed on day 18 to examine the number of implantation sites and live fetuses.<BR>In fresh medium, 185 of 228 zygotes (81%) were developed to blastocyst stage. The proportion of blastocysts developed from zygotes cultured in preserved media was not significantly different from that obtained in fresh medium (76-85%, in table 2).<BR>The proportions of implantation sites (58 to 65%) and live fetuses (38 to 51%) after transfer of blastocysts developed in preserved media were not also significantly different from those (67% and 46%) obtainded in fresh medium (Table 3).<BR>The present study clearly demonstrated that the chemically defined culture medium, M16, could be preserved at least for 90 days in refrigerator without any detrimental effect on
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